Publication details

Competitive Upconversion-Linked Immunosorbent Assay for the Sensitive Detection of Diclofenac

Authors

HLAVÁČEK Antonín FARKA Zdeněk HUEBNER Maria HORŇÁKOVÁ Veronika NĚMEČEK Daniel NIESSNER Reinhard SKLÁDAL Petr KNOPP Dietmar GORRIS Hans Heiner

Year of publication 2016
Type Article in Periodical
Magazine / Source Analytical Chemistry
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://pubs.acs.org/doi/abs/10.1021/acs.analchem.6b01083
Doi http://dx.doi.org/10.1021/acs.analchem.6b01083
Field Analytic chemistry
Keywords antibody; bioconjugation; diclofenac; immunoassay; pharmaceutical micropollutant; photon-upconversion
Description Photon-upconverting nanoparticles (UCNPs) emit light of shorter wavelength under near-infrared excitation and thus avoid optical background interference. We have exploited this unique photophysical feature to establish a sensitive competitive immunoassay for the detection of the pharmaceutical micropollutant diclofenac (DCF) in water. The so-called upconversion-linked immunosorbent assay (ULISA) was critically dependent on the design of the upconversion luminescent detection label. Silica-coated UCNPs (50 nm in diameter) exposing carboxyl groups on the surface were conjugated to a secondary anti-IgG antibody. We investigated the structure and monodispersity of the nanoconjugates in detail. Using a highly affine anti-DCF primary antibody, the optimized ULISA reached a detection limit of 0.05 ng DCF per mL. This performance came close to a conventional enzyme-linked immunosorbent assay (ELISA) without the need for an enzyme-mediated signal amplification step. The ULISA was further employed for analyzing drinking and surface water samples. The results were consistent with a conventional ELISA as well as liquid chromatography–mass spectrometry (LC–MS).
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